What plating conditions would you recommend for Nuff cells?

The recommended seeding density is 20,000 to 30,000 cells per cm2. Nevertheless, culture conditions might be adapted according to the iPS cells used.

NuFF cells should be plated 24 hours prior to plating the pluripotent cells, and can be used for 7–10 days.

If the cells are too sparse, they may not maintain the pluripotent cells without differentiation, and the pluripotent cells may not attach well

If they are too dense, they may detach from the plate, and the culture will be lost

What medium should I use to grow treated NuFF cells ?

NuFFs are expanded in DMEM containing 2 mM glutamine + 15% FBS. Treated NuFF cells should be thawed and plated into the same medium. When using the Newborn Human Foreskin Fibroblasts as a feeder layer, the Stem cell medium can be used when the “stem cells” are plated on the feeders.

How many times can NuFF cells be expanded?

NuFF cells can be passaged to P11 and used to culture pluripotent stem cells.

Cells can be expanded much further, but no tests have been performed above passage 11.

Is it possible to have access to untreated Human Newborn Foreskin Fibroblasts?

Functionally validated by Stemgent-Asterand to support iPS cell colonies from BJ human fibroblasts using the Stemgent’s mRNA Reprogramming Kit

While a range of mitotically inactivated Human fibroblasts can be used as a support layer for pluripotent cell culture, not all will support mRNA reprogramming of patient dermal fibroblasts. NuFF-RQ™ are 100% guaranteed for Stemgent’s RNA cell reprogramming. Browse through Stemgent’s user manual on Stemgent-Asterand’s website for further details.

Various techniques supporting cell reprogramming and iPS cells are now available (ex. small molecules, stabilized FGF2, Activin-A or EGF Stembeads growth factors…). At the end of this post, why not share your experience regarding the way you have been able to boost the efficiency of your cell reprogramming strategies.

You can also leave questions to cell biology experts who’ll get back to you to suggest appropriate solutions and tips!